-NRLF. 


o 


LIBRARY 

OF  THE 

UNIVERSITY  OF  CALIFORNIA. 


BIOLOGY 

LIBRARY 

G 


RECEIVED    BY   EXCHANGE 
Class 


STUDIES  OF  THE  INFLUENCE  OF  VARIOUS 
DIETARY  CONDITIONS  ON  PHYSIO- 
LOGICAL RESISTANCE. 

II.     THE    INFLUENCE   OF   DIFFERENT    PRO- 
PORTIONS OF  PROTEIN  IN  THE  FOOD 
ON  THE  PARTITION  OF  URINARY 
NITROGEN  AFTER  DOSAGE 
WITH    POTASSIUM 
CYANID. 


BY 

WllXIAM  H.  WELKER,  A.  C., 
In  collaboration  with  NORMAN  E.  DITMAN,  M.D. 


DISSERTATION 

Submitted  in  partial  fulfilment  of  the  requirements  for  the  degree  of  Doctor 

of  Philosophy,  in  the  Faculty  of  Pure  Science, 

Columbia    University. 

NEW  YORK  CITY 
1909 


STUDIES  OF  THE  INFLUENCE   OF  VARIOUS 
DIETARY  CONDITIONS  ON  PHYSIO- 
LOGICAL RESISTANCE. 

II.     THE    INFLUENCE   OF    DIFFERENT    PRO- 
PORTIONS OF  PROTEIN  IN  THE  FOOD 
ON  THE  PARTITION  OF  URINARY 
NITROGEN  AFTER  DOSAGE 
WITH    POTASSIUM 
CYANID. 


BY 

WILLIAM  H.  WELKER,   A.  C., 
In  collaboration  with  NORMAN  E.  DITMAN,  M.D. 


DISSERTATION 

Submitted  in  partial  fulfilment  of  the  requirements  for  the  degree  of  Doctor 

of  Philosophy,  in  the  Faculty  of  Pure  Science, 

Columbia    University. 

NEW  YORK  CITY 
1909 


BIOLOGY 

Ltr      ••: 


PREFACE 

This  dissertation  describes  experiments  which,  in  point  of 
inauguration,  were  the  second  group  in  a  series  of  studies  begun 
in  this  laboratory  in  1905  by  Dr.  Gies  with  Dr.  N.  B.  Foster's 
co-operation.  The  results  of  the  first  study  of  the  series  have 
lately  been  published  in  a  preliminary  communication  by  Foster: 
Proceedings  of  the  Society  for  Experimental  Biology  and  Medicine, 
1909,  vi,  p.  61. 

This  dissertation  contains  data  that  will  be  included,  for  the  most 
part,  in  a  paper  on  certain  effects  of  deficient  oxidation,  to  be 
published  from  this  laboratory  by  Ditman  and  Welker,  and  which 
will  appear  in  the  New  York  Medical  Journal  on  or  about  May  15, 
1909-  .  . 

•  •;  .it  is  Djy(pje£':ii:ctention  to  continue  the  study  of  the  influence 

physiological  resistance. 


of  *vario*us"  dietaj-y^  conditions  on 


WILLIAM  H.  WELKER. 


Laboratory  of  Biological  Chemistry  of  Columbia  University, 

College  of  Physicians  and  Surgeons, 

January  15,  1909. 


Reprinted   from   the  Neiv   York  Medical  Journal  for 
July  n,  1908. 


ON     SOME    BIOCHEMICAL    AND    ANATOMICAL 
CHANGES     INDUCED     IN     DOGS     BY 

POTASSIUM  CYANIDE. 

BY  WILLIAM  H.  WELKER,  A.  C,  AND  NORMAN  E. 

DITMAN,  M.  D., 

New  York, 

{From   the   Laboratories   of  Biological   Chemistry   and  Pathology   of 

Columbia   University,   at  the   College  of  Physicians 

and  Surgeons,  New  York,) 

I. — Introduction. 

In  connection  with  an  investigation  that  was  lately 
conducted  by  us  in  these  laboratories,  on  diminished 
oxidation  under  certain  pathological  conditions,  it 
was  desirable  to  ascertain  the  effects  of  such  dimin- 
ished oxidation  on  elimination  of  various  nitrogen 
containing  products  in  the  urine.  Potassium  cyanide, 
which  induces  diminished  oxidation,  was  selected 
for  the  purpose,  and  the  six  experiments  described 
in  this  paper  were  accordingly  carried  out.  These 
experiments  were  also  intended  to  furnish  funda- 
mental data  for  investigations  of  the  influence  of 
guanidin,  methyl  guanidin,  creatin,  and  creatinin 
under  conditions  of  diminished  oxidation.  We  have 
carried  out  preliminary  experiments  along  these 
lines,  and  hope  to  conduct  them  before  long  to  com- 
pletion. 

It  had  been  our  intention  to  carry  out  several  ad- 
ditional experiments  before  making  our  data  public, 
but  the  recent  appearance  of  a  paper  on  this  partic- 
ular subject,  which  presented  results  (of  only  a  few 
experiments)  that  are  not  in  harmony  with  those 
obtained  in  this  study,  induced  us  to  publish  at 

Copyright,   1908,  by  A.   R.  Elliott  Publishing  Company. 


218494 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

once  the  observations  we  have  made,  especially  as 
it  will  be  some  time  before  we  shall  be  able  to  re- 
sume the  work  in  this  connection. 

2. — Conduct  of  the  Experiments. 

Animals  and  environment. — These  experiments 
were  conducted  on  six  dogs  by  the  general  meta- 
bolism methods  used  in  this  laboratory.1  Each  ani- 
mal was  apparently  healthy  and  normal  in  all 
respects  at  the  beginning  of  the  experiment  in  which 
it  was  used.  The  form  of  the  cage2  in  which  the 
dogs  were  confined  was  very  well  adapted  to  the 
collection  of  the  excreta  and  allowed  rapid  substitu- 
tion of  urine  receivers,  when  vomiting  very  closely 
followed  the  elimination  of  urine  or  vice  versa.8 
The  cage  as  well  as  the  room  was  well  ventilated. 
The  animals  were  thoroughly  accustomed  to  their 
surroundings  and  apparently  contented  before  they 
were  subjected  to  experimental  treatment. 

Food. — The  daily  diet  consisted  of  hashed  lean 
meat,  cracker  meal,  lard,  bone  ash  and  water.  The 
raw  meat  was  preserved  by  refrigeration,  according 
to  the  usual  method  in  this  laboratory.4  The  bone 
ash  was  mixed  with  the  food  to  give  the  faeces  a 
desirable  consistency.5 

Periods,  weights. — In  the  records  of  the  experi- 
ments, each  day  ended  at  n  a.  m.  Just  before  that 
time  the  animal  was  weighed  and  the  weight 
recorded  for  the  day.  At  that  time  also  the  excreta 
were  removed  from  the  cage  and  urine  receiver. 
The  food  was  then  given. 

Injection  of  potassium  cyanide. — The  potassium 

1Mead  and  Gies:  American  Journal  of  Physiology,  v,  p.  106, 
1901;  also  Gies  and  Collaborators:  Biochemical  Researches,  i,  p. 
419  (Reprint  No.  21),  1903. 

2Gies:  American  Journal  of  Physiology,  xiv,  p.  403,   1905. 

8Berg  and  Welker:  Journal  •/  Biological  Chemistry,  i,  p.  410, 
1906;  Welker:  American  Journal  of  Physiology,  xx,  p.  358,  1907. 

*Gies:  American  Journal  of  Physiology ,  v,  p.  235,  1901;  Gies  and 
Collaborators:  Biochemical  Researches,  i,  p.  69  (Reprint  No.  i), 
1903. 

5Steel  and  Gies:  American  Journal  of  Physiology,  xx,  p.  343,  1907. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

cyanide  used  in  these  experiments  was  a  sample  of 
Kahlbaum's  purest  ("K").  Whenever  used,  the 
cyanide  was  rapidly  weighed  and  a  I  per  cent,  solu- 
tion prepared.  This  solution  was  protected  from 
direct  light  when  not  in  use.  Only  fresh  solutions 
were  employed.  The  injections  were  made  sub- 
cutaneously  in  the  lumbar  region.  The  animals 
were  in  no  instance  put  deeply  under  the  influence 
of  the  cyanide  by  the  amounts  injected. 

Analytic  methods. — No  analyses  were  made  of 
the  food  materials  used  in  these  experiments,  but 
great  care  was  taken  to  secure  a  sufficient  supply  of 
each  of  the  various  ingredients  to  last  throughout  an 
entire  experiment.  Uniformity  of  diet  from  begin- 
ning to  end  of  each  experiment  was  thus  assured. 

The  amount  of  ammonia  in  the  urine  was  deter- 
mined daily  in  all  experiments  except  the  fifth,  in 
which  experiment  the  dog  was  on  a  relatively  high 
plans  of  protein  nutrition.  In  the  case  of  that  ani- 
mal, it  was  found  by  experiment  that  the  urine 
which  was  preserved  with  thymol,  showed  little  ten- 
dency to  undergo  change  of  ammonia  content  even 
after  several  days,  and  ammonia  was  accordingly 
determined  in  combined  daily  urines.  The  remain- 
ing determinations  were  made  on  "period  urines." 
These  period  urines  were  prepared  by  combining 
aliquot  fractions  of  the  daily  volumes.  In  the  case 
of  the  first  dog  the  volumes  of  the  "period  urines" 
were  so  low  that  they  were  diluted  with  equal  quan- 
tities of  water,  in  order  that  the  usual  amounts  could 
be  used  for  the  various  determinations.  The  meth- 
od used  for  the  determination  of  ammonia  in  the 
urine  was  Folin's.8 

Total  nitrogen  was  determined  by  the  use  of  the 
Kjeldahl  method.  The  urine  was  decomposed  by 
means  of  concentrated  sulphuric  acid  and  small 
quantities  of  copper  sulphate,  a  method  long  in  satis- 

6Folin:  Zeitschrift  fur  physiologische  Chemie,  xxxii,  p.   161,   1902. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

factory  use  in  this  laboratory.  The  digestion  was 
continued  for  thirty  to  forty-five  minutes  after  the 
mixture  had  assumed  a  true  greenish  tint.  On  cool- 
ing, in  each  case  the  mixture  was  colorless. 

The  alloxuric  nitrogen  was  determined  by  pre- 
liminary precipitation  according  to  the  Salkowski7 
method,  followed  by  treatment  of  the  washed  pre- 
cipitate, together  with  the  nitrogen-free  filter  paper, 
by  the  A rn stein8  method  for  the  removal  of  ammonia 
nitrogen,  i.  e.,  by  adding  water  and  magnesium  ox- 
ide to  the  paper  and  precipitate,  and  boiling  the  mix- 
ture. To  facilitate  matters  this  was  done  in  a  large 
Kjeldahl  flask.  After  all  the  ammonia  had  been 
driven  off,  the  residual  nitrogen  was  estimated  by 
the  usual  Kjeldahl  method. 

Urea  was  determined  by  the  Folin9  method ;  allan- 
toin  by  the  Loewy10  method ;  uric  acid  by  the  Folin- 
Shaffer11  method ;  creatinin  by  the  Folin12  method. 

Summaries  of  observations  and  results  in  the  experi- 
ments with  animals  on  medium  planes  of  pro- 
tein nutrition  (Tables  I — IV.). 

Table  L— First  Dog. 

Preparatory  period. — The  daily  diet  of  this  ani- 
mal consisted  of  meat,  97  grammes  ;13  cracker  meal, 
26  grammes ;  lard,  20  grammes ;  bone  ash,  10 
grammes ;  water,  225  c.c.  The  initial  weight  of  the 
animal  was  6.5  kilos.  The  preparatory  period  con- 
tinued for  seventeen  days. 

7Salkowski:  PHuger's  Archiv,  Ixix,  p.  268,   1897. 

8Arnstein:  Zeitschrift  fur  physiologische  Cheinie,  xxiii,  p.  417, 
1897;  Centralblatt  fur  die  medizinischen  Wissenschaften,  xxxvi,  p. 
257,  1898. 

9Folin:  Zeitschrift  fur  physiologische  Chemie,  xxxii,  p.  504,  1901; 
xxxvi,  p.  333,  1902;  xxxvii,  p.  548,  1903. 

10Loewy:  Archiv  fur  experimentelle  Pathologic  und  Pharmakologie. 


44,   p.    19,    1900. 

"Folin  and  Shaffer:  Zeitschrift  fur  physiologische  Chemie,  xxxii, 
p.  552,  1901. 

12Folin:  Ibidem,  xli,  p.  223,    1904. 

1:5 About  15  grammes  per  kilo;  approximately  0.53  gramme  of  ni- 
trogen per  kilo. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 
Table  I. — First  Dog. 

FIRST   PERIOD. — NORMAL   CONDITIONS,   JULY    5   TO    13,    1907. 

•8 


5-94 
5-94 
5-94 
5-92 
5-92 
5-96 
5-95 
5-93 
5-97 


170 
130 
i85 

95 
170 


21 
20 
22 
21 
24 
28 
26 
19 
26 


..2  53 


Acid. 

Faintly   acid. 
Acid. 

Amphoteric. 
Acid. 


_c  c  £  g 

Slf  ft 

0.1196 

0.0912 
0.1582 
0.1034 
0.0926 
0.1165 
0.0885 
0.1276 


^    H 


3*.» 


SECOND   PERIOD. — EFFECTS   OF   POTASSIUM    CYANIDE    (SMALL   DOSES), 
JULY   14  TO  25,   1907. 


II.... 

Acid 

12  .  .  .  . 

5  91 

I  6O 

ll 

o  0986 

20 

e  og 

u 

14  — 

...      5-95 

...       5-93 
•  •  .       5.96 

no 
90 

1  20 

28 
29 
28 

Faintly  alkaline. 
Amphoteric 

0.1127 
0.0924 
0.1233 

15 
26 
12 

17  — 

!g  

...      5-97 

e  qi 

130 

20 

Acid 

o.i353 

21 
2O 

19  .  .  .  . 

.  .  .       591 

s 

22 

Amphoteric. 

O.I2I  I 

10, 

20 

26 

Neutral 

I  e 

21.  . 

5.94 

14  ; 

20 

O.IOI3 

II 

THIRD  PERIOD.U — FURTHER  EFFECTS   OF  POTASSIUM   CYANIDE    (RELA- 
TIVELY LARGE  DOSES),  JULY  26  TO  29,   1907. 


23 

40 
1  60 

Acid 

0.0351 

18 

«33 

24.  . 
25  .  . 

90 
170 

29 

Faintlv  alkaline. 
Acid 

0.0629 
o  1388 

8 

25 

»3 

"3 

"The  urine  of  this  period  contained  a  small  quantity  of  the 
liquid  portion  of  the  vomit. 

15Vomited  sixteen  minutes  after  injection.  A  few  c.  c.  of  the 
liquid  part  of  the  vomit  were  unavoidably  mixed  with  some  of  the 
urine  in  the  receiver.  The  solid  vomitus  was  eaten  again,  shortly 
afterward  when  offered  to  the  animal. 

16 Vomited  twenty  minutes  after   injection.     Ate  all  vomitus   later. 

"Vomited   seven    minutes    after   injection.     Ate    all    vomitus    later. 

"Vomited  twenty  minutes  after  injection.     Ate   all  vomitus   later. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 
Table  I. — First  Dog — Continued. 

FOURTH    PERIOD. — FURTHER    EFFECTS    OF    POTASSIUM    CYANIDE    (IN- 
CREASED DOSES),  JULY  30  TO  AUGUST  6,  1907. 


i° 


di 


•5  ^ 


C 
re 

"       I 

C        p 

.2  oa 

»!s  S> 


26  

27  .  .  . 

5-86 
S  81 

1  60 

I  2O 

22 

26 

Faintly    acid. 
Acid. 

o.i  1  66 
0.0723 

13 

3.5 

19-    - 

28 

S.8e 

60 

48 

0.0940 

IO 

2035 

2Q 

S8S 

I  IO 

tt 

o.  1  024 

20 

o-  0 

3c 

30  

n 
5.85 

5.85 

120 

27 
c  e 

Alkaline. 
Faintly    alkanie. 

0.0906 
o.  1  043 

18 

1  0 

•o 

32  

145 

OO 
24 

Alkaline. 

o.i  390 

25 

3 

33.  . 

s'.so 

I  SO 

20 

.Neutral. 

O.I  023 

30 

22o"e 

FIFTH  PERIOD. — AFTER  PERIOD,  AUGUST   7  TO   16,    1907. 


•je 

3-"0 

e  80 

126 

I§ 

ii 

o  081  5 

23 

si:::::  . 

5  80 

1  *5 

•3C 

ii 

o  1  534 

20          .  . 

37  •  •  ' 

5-77 

190 

jg 

(i 

o.  1343 

23          .  . 

38 

e   80 

26 

ii 

on.  .  . 

c  80 

I  SO 

26 

0              .  . 

40  

5.80 

no 

23 

Alkaline. 

o  0633 

22 

5  80 

42  

43  

5-73 
e  74 

115 

I2O 

29 

Amphoteric. 
Alkaline 

0.0862 

12 
I  c 

"Vomited  repeatedly,  beginning  eighteen  minutes  after  injection, 
ending  fifteen  minutes  afterward.  Ate  all  vomitus  later. 

20Vomited  eighteen,  and  again  thirty-four  minutes  after  injection. 
Ate  all  vomitus  later. 

21Vomited  several  times  thirty  minutes  after  injection.  Ate  all 
vomitus  shortly  afterward. 

22Diarrhcea  occurred  near  the  end  of  the  day. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


Table  III.— Second  Dog. 

Preparatory  period. — The  initial  weight  of  this 
animal  was  8.87  kilos.  The  preparatory  period  con- 
tinued for  13  days.  The  daily  diet  consisted  of  the 
following:  Meat,  135  grammes;23  cracker  meal,  36 
grammes ;  lard,  27  grammes ;  bone  ash,  10  grammes ; 
water,  315  c.c. 

FIRST   PERIOD. — NORMAL   CONDITIONS,    OCTOBER  21   TO  29,    1907. 


At 


.2  -•>  <n 


E    -cgi 


S  «-*•- 
£  ofc.| 
so fc  &s 


8.36 
8-33 
8-34 
8-35 
8.35 
8.30 
8.32 
8.28 
8-35 


405 

359 
330 
290 
330 
375 
305 
420 
382 


Acid. 


0.1997 
0.2281 
0.1844 

0.1573 

0.1681 
0.2034 
0.1537 
0.1772 
0.1716 


SECOND  PERIOD. — EFFECTS  OF  POTASSIUM   CYANIDE,   OCTOBER 
NOVEMBER  6,   1907. 


30  TO 


8.28 
8.28 
8.29 
8.26 
8.30 
8.25 
8.30 
8.30 


309 
360 
365 
365 
320 
425 
335 
340 


Acid. 


0.1913 
0.1779 
0.1640 
0.1620 
0.1245 
0.1723 
0.1652 
0.1932 


10 

o 

40 
16 
16 
28 
27 


o 

2 
2.25 

2-5 

2-75 

3 


THIRD   PERIOD. — AFTER   PERIOD,   NOVEMBER    7   TO    12,    1907. 


X8  

.  8  ^6 

284 
387 

Ifi 
& 

16   Ac 
14 
15 
15 
15 
18 

id.   0.1385 
0.1526 
0.1595 
0.1690 
0.1195 
0.1130 

id 

31 
o 
35 

11 

19  

g.^o 

8  ?•? 

8  17 

22  

8  \2 

23  

8  is 

28 About   is   grammes  per  kilo;    approximately  0.53   gramme  of   ni- 
trogen per  kilo. 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

Discussion  of  the  chemical  results  of  the  experi- 
ments with  animals  on  medium  planes  of  pro- 
tein nutrition  (first  and  second  dogs,  Tables 
I-IV}. 

The  injection  of  small  doses  of  cyanide  in  the  case 
of  the  first  dog  (Period  II,  Tables'l  and  II)  caused 
a  decrease  in  the  average  daily  excretion  of  total 
nitrogen  in  the  urine.  There  was  a  corresponding 
decrease  in  the  amounts  of  all  the  various  forms  of 
nitrogen  named  in  the  Tables  (I  and  II),  except 
that  of  the  purin  bases,  which  was  increased. 

The  analytic  results  for  the  third  period  (Tables 
I  and  II),  in  which  the  daily  doses  of  the  cyanide 
were  3  nigs,  per  kilo,  are  not  uniform  enough  to  tell 
a  definite  or  concordant  story.  The  period  was  per- 
haps comparatively  too  short  for  the  full  registra- 
tion of  the  effects  that  may  have  been  induced  by 
the  cyanide.  Then,  too,  contamination  of  the  urine 
with  vomit  may  have  introduced  some  of  the  ap- 
parent uncertainties  of  significance  in  the  results. 

In  the  fourth  period,  during  which  the  doses  of 
cyanide  were  3^  mgs.  per  kilo,  the  average  daily  ex- 
cretion of  total  nitrogen  was  less  than  that  of  the 
first  or  normal  period.  This  was  also  true  of  the 
various  other  forms  of  nitrogen,  except  the  "unde- 
termined nitrogen,"  which  increased  in  amount  in 
this  period.  The  daily  amount  of  nitrogen  in  purin 
bases,  which  had  risen  considerably  in  the  first  dos- 
age period  and  fallen  in  the  next,  was  so  small  as  to 
be  practically  nil  in  the  third  dosage  period. 

If  cyanide  has  a  tendency,  as  claimed  by  previous 
observers,  to  increase  nitrogenous  elimination,  we 
should  expect  to  find  an  exhibition  of  this  effect  in 
the  fourth  period,  during  which  comparatively  large 
doses  of  cyanide  were  given,  unless  the  cyanide 
treatment  in  the  preceding  periods  had  altered  the 
basis  for  further  comparisons.  As  indicated  above, 
however,  decrease  rather  than  increase  was  the  rule, 
except  in  the  third  period.  In  the  after  period  slight 

10 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

increases  in  most  of  the  figures  in  the  tables  are  to 
be  noted. 

During  the  period  of  cyanide  injection  in  the  ex- 
periment on  the  second  dog,  the  average  daily  quan- 
tities of  total  nitrogen,  of  the  nitrogen  of  urea,  of 
that  of  purin  bases,  and  of  creatinin,  were  increased, 
while  the  quantities  of  nitrogen  of  ammonia,  allan- 
toin,  uric  acid,  and  the  undetermined  fraction  were 
decreased. 

The  average  dose  of  cyanide  for  this  dog  was 
only  about  2  mg.  per  kilo.  No  vomiting  ever  oc- 
curred, and  from  all  outward  appearances  the  effects 
of  the  treatment  were  much  less  marked  than  in  the 
case  of  the  first.  In  spite  of  this  fact,  however,  the 
total  nitrogen  elimination  from  this  dog  increased, 
but  from  the  first  dog  decreased,  with  cyanide  treat- 
ment. 

Histological  data — First  and  second  dogs. 

Microscopical  examination  of  the  viscera  of  these 
dogs  showed  the  following: 

First  dog.  Kidney — The  uriniferous  tubules 
contained  a  large  amount  of  degenerated  cellular 
matter,  which  was  granular  in  appearance.  The 
epithelium  of  the  convoluted  tubules  was  slightly 
swollen  and  irregular  in  outline,  with  a  few  areas 
of  degeneration.  There  was  slight  proliferation  of 
connective  tissue  along  the  course  of  the  small 
bloodvessels. 

Liver — This  was  the  seat  of  a  degenerative  pro- 
cess of  irregular  intensity  in  its  distribution,  with 
no  definite  relation  to  the  periphery  of  the  lobule 
or  the  central  vein.  The  degeneration  was  albu- 
minous in  type  and  moderate  in  intensity — many 
liver  cells  being  simply  the  seat  of  extreme  vacuoli- 
zation.  The  nuclei  were  unchanged. 

Heart — Normal.  Spleen — Normal.  Pancreas — 
Normal. 

n 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

Second  dog.  Kidney — There  was  moderate  al- 
buminous degeneration  of  the  tubular  epithelium, 
which  was  most  marked  in  the  convoluted  tubules. 
Some  contained  casts.  There  were  numerous  areas 
of  small  round  cell  proliferation  along  the  blood- 
vessels and  about  the  glomeruli.  A  few  of  these 
areas  were  infiltrated  with  pus  cells. 

Liver — There  was  well  marked  albuminous  de- 
generation, uniform  in  distribution,  with  marked 
vacuolization  of  the  cells.  There  was  no  conges- 
tion. The  nuclei  were  unchanged. 

Heart — Normal.  Spleen — Normal.  Pancreas- 
Normal. 

Summaries  of  observations  and  results  in  the  ex- 
periments with  animals  on  low  planes  of  protein 
nutrition  (Tables  V-X). 

Table  V.— Third  Dog. 
Preparatory  Period. 

Table  showing  the  decreasing  quantities  of  meat 
fed  the  animal  daily  in  bringing  it  gradually  down 
to  a  low  plane  of  protein  nutrition.  (Initial  daily 
diet:  Meat,  96  grammes;24  cracker  meal,  32 
grammes  ;  lard,  24  grammes  ;  bone  ash,  8  grammes  ; 
water,  280  c.c.) 


Date. 

Body  weight, 
kilos. 
8.00 

Weight  of  meat, 
grammes. 
96 

8.00 

96 

7.93 

96 

7.Q2 

96 

8.OO 

qi 

8.00 

86 

1       f   ' 

7-92 

81 

7.88 

76 

uly  18 

.    ..            7  90 

71 

....            7-9O 

66 

7.92 

61 

7.86 

56 

uly  22  

7.82 

25S6 

24 About  12  grammes  per  kilo;  approximately  0.42  gramme  of  ni- 
trogen per  kilo. 

~>5About  7  grammes  per  kilo;  approximately  0.25  gramme  01  ni- 
trogen per  kilo. 

12 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

The  only  ingredient  in  the  diet  that  was  subject 
to  quantitative  change  during  the  preparatory  period 
was  the  meat.  Two  days  previous  to  the  beginning 
of  the  fore  period  and  from  then  on  to  the  end  of 
the  experiment,  the  quantity  of  meat  fed  daily  was 
56  grammes.  The  other  ingredients  were  as  fol- 
lows :  Cracker  meal,  32  grammes  ;  lard,  24  grammes ; 
bone  ash,  8  grammes ;  water,  280  c.c. 

Table  VI.— Third  Dog. 

FIRST  PERIOD.— NORMAL  CONDITIONS,  JULY  23  TO  28,   1907. 

4 


p 


7.81    290    ii      Acid.      0.1135    10 


2 7.78  260  ii 

3 7-74  290  10 

4 7-82  195  13 

5 7.81  245  TO 

6 7-73  340  10 


O.I2II  27 

O.II9I  II 

O.I  1 22  8 

O.0973  22 

0.0978  18 


SECOND    PERIOD. — EFFECTS    OF    POTASSIUM     CYANIDE,    JULY    29    TO 
AUGUST    5,    1907- 

7 7.67         300         12               Acid.  0.1413  32 

8 7-65         303         10  0.2946  ..  i 

9 7-65         255         10  0.1432  31  1.5 

10 7.65         223         ii  0.5027  17  1.5 

ii 7.64         295         ii            Alkaline.  0.4606  20  2 

12 7.65         255         ii  0.2871  23  2.5 

13 7-55          290          12                Acid.  0.1986  39      -63 

14 7.55         270         ii           Alkaline.  0.1960  9  2.5 

THIRD  PERIOD. — AFTER  PERIOD,  AUGUST  6  TO   13,  1907- 


16.  .  . 

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« 

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205 

« 

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1  1 

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O.I222 

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20  ... 

21  C 

°s 

O.O5OI 

6 

21  ... 

—    7.70 

290 

ii 

Acid. 

0.1803 

JO 

20This  quanitity  of  potassium  cyanide  caused  vomiting  about  one- 
half  hour  after  injection,  also  mild  diarrhoea,  followed  by  marked 
prostration.  Later  the  animal  appeared  to  be  normal  and  ate  all 
of  the  vomitus. 

13 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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27The  differences  between  the  results  of  the  determinations  of  the 
alloxuric  nitrogen  and  the  uric  acid  were  so  slight  as  to  ^warrant 
the  conclusion  that  the  uric  acid  nitrogen  was  the  only  purin  nitro- 
gen eliminated  in  quantities  sufficiently  large  for  consideration. 

14 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

Table  VIII.— Fourth  Dog. 

Preparatory   Period,   October    n    to   November   2, 
1907. 

Table  showing  the  diminishing  quantities  of  meat 
fed  to  the  animal  in  bringing  it  gradually  down  to 
the  low  nitrogen  plane.  (Initial  daily  diet:  Meat, 
1 60  grammes;  cracker  meal,  40  grammes;  lard,  30 
grammes;  bone  ash,  10  grammes;  water,  350  c.c.) 


I  4 

I  |  i  j 

I              ii  II  I  li  II 

Q                                    K3  3&  tt  CSS  S  S> 

Oct.    ii 9.48  160             Oct.  22 8.76  105 

Oct.  12 9.26  28iss      Oct.  23 8.71  29ioo 

Oct.  13 9.05  150      Oct.  24 8.64  95 

Oct.  14 9.10  145      Oct.  25 8.75  90 

Oct.  15 9.06  140     Oct.  26 8.61  85 

Oct.  16 8.90  135      Oct.  27 8.58  80 

Oct.  17 8.89  130  Oct.  28 75 

Oct.  18 8.87  125      Oct.  29 8.53  70 

Oct.  19 8.85  120     Oct.  30 8.57  70 

Oct.  20 8.73  115      Oct.  31 8.59  70 

Oct.  21 8.79  no     Nov.  i 8.39  70 

Nov.  2 8.52  807O 


The  meat  was  the  only  ingredient  that  was  sub- 
ject to  quantitative  change  during  the  preparatory 
period.  At  the  beginning  of  the  fore  period,  and 
throughout  the  whole  experiment  thereafter,  70 
grammes  of  meat  were  given  daily  with  the  fol- 
lowing amounts  of  the  remaining  ingredients  of  the 
diet :  Cracker  meal,  40  grammes  ;  lard,  30  grammes  ; 
bone  ash,  10  grammes ;  water,  350  c.c. 


28  About  17  grammes  per  kilo;  approximately  0.6  gramme  of 
nitrogen  per  kilo. 

29About  11.5  grammes  per  kilo;  approximately  0.4  gramme  of 
nitrogen  per  kilo. 

30 About  8.25  grammes  per  kilo;  approximately  0.3  gramme  of 
nitrogen  per  kilo. 

IS 


Wclker  and  Ditman:  Biochemical  and  Anatomical  Changes. 
Table  IX.— Fourth  Dog. 

FIRST  PERIOD. — NORMAL  CONDITIONS,  NOVEMBER  3  TO  7,  1907- 


I  &    c    i 

I         1        f  .,  f.         ig|  tgljl 

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£                      U.2          'C'o0  '^{£.0  *CgS                   'Cg-Sj 

O                    P5^2         P>cj  Pco  «  Prtvs                P<5B6 

i 8.41         423  ii  Acid.               0.2085         19 

2 8.52         200  09     Faintly  alkaline.     0.0412         28 

3 8.48         360  ii  Acid.               0.1637         14 

4 8.39         394  10  0.1545          19 

S 8.33         386  12  0.1945          :5 

SECOND    PERIOD. — EFFECTS    OF    POTASSIUM    CYANIDE,     NOVEMBER    8 

TO  13,  1907- 

6 8.29         300  ii  Acid.               0.1446         29        i 

7 8.28         335          10  0.1233          ii         i-5 

8 8.25         335          ii  0.1360  30        2. 

9 8.20         254  07  0.0654          18     312.5 

10 8.29         311          13  0.1545          IS     322 

II 8.23             366             II  O.I905             20       832 

THIRD  PERIOD. — AFTER  PERIOD,  NOVEMBER   14  TO  20,  1907. 


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Discussion  of  the  chemical  results  of  the  experi- 
ments with  animals  on  low  planes  of  protein 
nutrition  (third  and  fourth  dogs,  Tables  V-X). 
The  average  daily   total  urinary  elimination  of 
nitrogen  in  these  two  experiments  failed  to  show  a 
concordant  tendency.     In  the  case  of  the  third  dog 
(Tables  VI-VII)  there  was  a  slight  increase  dur- 
ing the  cyanide  period,  while,  on  the  other  hand, 
there  was  a  decrease  in  the  case  of  the  fourth  dog 
(Table  IX-X)   during   the    cyanide  period.      The 

81Vomited  repeatedly  during  the  first  half  hour  after  injection. 
Refused  all  offers  of  vomitus,  but  ate  the  same  with  the  regular 
meal  on  the  following  day. 

32 Appetite  diminished.  Vomited  within  twenty  minutes  after  in- 
jection. Ate  all  of  vomitus  later. 

33Vomited  about  fifteen  minutes  after  injection.  Ate  the  vomitus 
with  the  meal  on  the  following  day. 

16 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

nitrogen  of  ammonia  was  markedly  increased  in 
the  case  of  the  third  dog  during  the  cyanide  period 
(Tables  VI-VII),  but  was  diminished  slightly  in 
the  experiment  on  the  fourth  dog  (Tables  IX-X). 
The  average  daily  elimination  of  urea  was  very 
slightly  increased  during  the  cyanide  period  in  the 
experiment  on  the  third  dog,  but  in  terms  of  per 
cent,  of  the  total  nitrogen  it  decreased  markedly. 
During  the  same  period  in  the  experiment  on  the 
fourth  dog,  however,  the  average  daily  amount  of 
urea  nitrogen,  as  well  as  its  per  cent,  of  the  total 
nitrogen,  decreased  decidedly.  The  other  notable 
result  of  these  two  experiments  was  the  sharp  in- 
crease in  the  average  daily  amount  of  the  undeter- 
mined nitrogen  in  each  experiment.  The  remaining 
results  of  these  two  experiments  are  diametrically 
opposed  to  each  other. 

Histological  data — Third  and  fourth  dogs. 

The  microscopical  examination  of  the  viscera  of 
these  dogs  showed  the  following: 

Third  dog.  Kidney — There  was  congestion  of 
the  cortical  area  and  slight  swelling  and  irregu- 
larity of  outline  of  the  cells  of  the  convoluted 
tubules. 

Liver — This  was  the  seat  of  a  diffuse  degenera- 
tive process,  uniform  in  intensity  in  all  parts  of  the 
liver  lobules.  The  nuclei  were  intact,  but  the  cyto- 
plasm was  the  seat  of  an  extreme  albuminous  de- 
generation, which  was  so  excessive  that  only  a  few 
areas  of  granular  cytoplasm  remained  and  the  out- 
lines only  of  the  cells  persisted.  There  were  ir- 
regular areas  of  congestion. 

Heart — Normal.  Spleen — Normal.  Pancreas — 
Normal. 

Fourth  dog.  Kidney — There  was  slight  swelling 
of  the  epithelial  cells  of  the  cortical  tubules,  with 
irregularity  of  outline.  There  was  a  little  exudate, 
Scattered  irregularly  through  the  cortex  were 
areas  of  small  round  cell  infiltration.  Most  of  these 

18 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

areas  contained  a  number  of  pus  cells.  They  oc- 
curred along  the  course  of  the  bloodvessels,  but  the 
glomeruli  were  not  involved. 

Liver — There  was  well  marked  albuminous  de- 
generation, uniform  in  distribution,  with  marked 
vacuolization  of  the  cells.  There  was  no  congestion. 
The  nuclei  were  normal. 

Heart — Normal.  Spleen — Normal.  Pancreas — 
Normal. 

Summaries  of  observations  and  results  in  the  ex- 
periments with  animals  on  high  planes  of  pro- 
tein nutrition  (Tables  XI-XVI}. 

Table  XL— Fifth  Dog. 

Preparatory  period. — Table  showing  the  increas- 
ing quantities  of  meat  fed  the  animal  in  raising  it 
to  the  high  nitrogen  plane. 

(The  initial  diet  of  this  animal  consisted  of  meat, 
1 20  grammes;  cracker  meal,  32  grammes;  lard,  24 
grammes;  bone  ash,  10  grammes;  water,  280  c.c.) 


Body  weight, 

Weight  of  meat, 

Date. 

kilos. 

grammes. 

July 
July 
July 

I:::::::::::::::: 

8.25 

8.21 

8.08 

3*120 

140 

160 

uly 

8  

7-92 

1  80 

uly 

9  

7.85 

200 

uly 

10  

7-92 

220 

uly 

it  

7-90 

240 

uly 

12  

7-94 

240 

ulv 

13  

8.00 

«240 

uly 

14  

8.00 

240 

ulv 

15  

8.02 

260 

July 
Tuly 

16  
17  

8.03 
8:09 

280 
300 

July 

18  

8.13 

320 

luly 

19  

8.17 

340 

luly 

20  

8.22 

360 

July 
July 

21  
22  

8.31 
8.37 

380 
3«380 

The  meat  in  the  daily  diet  was  increased  twenty 
grammes  each  day,  excepting  on  July  I2th,  I3th, 

34About  15  grammes  per  kilo;  approximately  0.53  gramme  of  ni- 
trogen per  kilo. 

SHAbout  30  grammes  per  kilo;  approximately  1.05  gramme  of  ni- 
trogen per  kilo. 

38 About  45  grammes  per  kilo;  approximately  1.58  gramme  of  ni- 
trogen per  kilo. 

19 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

and  I4th,  ending  with  July  2ist,  when  the  daily 
amount  was  380  grammes.  The  other  ingredients 
remained  constant  in  quantity  throughout  the  entire 
experiment. 

Table  XII. —Fifth  Dog. 

FIRST   PERIOD. — NORMAL  CONDITIONS,   JULY  23   TO   29,    1907. 


M!S 

8.37 
8.40 
8.50 
8.48 
8-53 
8.58 
8.63 


(. 

lid 


418 
450 
410 
400 

440 


Faintly    acid. 


SECOND    PERIOD. — EFFECTS    OF    POTASSIUM     CYANIDE,  JULY  30    TO 
AUGUST   2,    1907. 

8" 8.65              510             22                Neutral  53  s"i 

9 8.77             390             24            Amphoteric  43  89o 

io37 8.70             475             22                       "  44  4°i 

ii 8.63              165              35                   Acid  o  41o 

THIRD    PERIOD. — RECOVERY    FROM    EFFECTS    OF    POTASSIUM    CYANIDE, 
AUGUST  3  TO  7,   1907. 


44 

8  65 

9243 

JO 

36 

Acid 
Alkaline 

68 

14  .      . 

8  72 

ov 
26 

\cid 

8  80 

3OO45 

' 

16 

8  82 

C7040 

87The  small  doses  of  cyanide  injected  on  the  eighth  and  tenth 
days  (July  3oth  and  August  ist)  set  up  violent  disturbances  (ap- 
parently because  of  this  high  protein  diet),  so  that  it  was  impossible 
to  continue  the  cyanide  treatment  without  decreasing  the  protein  in 
the  food.  The  daily  quantity  of  meat  was  accordingly  lowered  to 
300  grammes  (about  34  grammes  per  kilo;  approximately  1.19 
gramme  of  nitrogen  per  kilo),  on  the  seventeenth  day  (August  8th), 
and  this  amount  was  fed  daily  thereafter  to  the  end  of  the  experi- 
ment. 

^No  apparent  effects. 

89Vomiting  occurred  some  time  after  feeding.  All  of  the  vomitus  was 
eaten  later. 

*°No  apparent  effects. 

^Vomiting  occurred:  all  offers  of  same  were  refused. 

^Vomited   during  the  night.      No  vomit   in   the   urine. 

43Urine  contained  fluid  vomit. 

^Diminished    appetite,    mild    diarrhoea. 

45Vomited  after  feeding.  Urine  contained  fluid  vomit.  Ate  solid 
vomitus  later. 

^Vomited.     Urine  contained  trace  of  fluid  vomit. 


20 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

FOURTH    PERIOD. — PREPARATORY   PERIOD  WITH    REDUCED  AMOUNT  OF 
MEAT  IN  THE  DAILY   DIET,  AUGUST  8  TO   16,   1907. 


I 

17 
18 
19 
20 
21 
22 
23 
24 
25 


' 


8.75 
8.79 
8.85 
8.88 
8.85 
8.88 
8.90 
8.89 
8.88 


420 
320 
410 
432 
475 
390 
380 
445 
465 


23 
30 
19 
23 
18 
25 
22 
22 
23 


Amphoteric 


Faintly   acid 


40 
51 
48 
40 
51 
37 
57 
37 
37 


FIFTH    PERIOD.—  EFFECTS      OF   POTASSIUM    CYANIDE,  AUGUST  17   TO 
18,    1907. 

8.73             345             21               Alkaline              22  47o.s 

8.88             440             21                                           60  o 


26 

27 


SIXTH   PERIOD.  —  NORMAL  CONDITIONS,  AUGUST    19   TO   23,    1907. 
28  ........     8.89  460  21  Faintly  acid          57 


29 
30 
31 
32 


8.90 
8.90 
8.93 
8.96 


450 
445 
455 
430 


Amphoteric 

Faintly   acid 
Amphoteric 


SEVENTH   PERIOD.  —  EFFECTS   OF   POTASSIUM    CYANIDE   IN    RELATIVELY 


SMALL  DOSES,  AUGUST  24  TO  30,  1907. 


33 
34 
35 
36 
37 
38 
39 


8.92 
9-00 
8.92 
8.94 
8.94 
9.00 
8.94 


485 
490 
525 
455 
320 
555 
495 


Alkaline 
Amphoteric 

Acid 
Faintly    acid 

Acid 

Faintly    acid 
Faintly   acid 


0.5 
0.5 
°-5 


i 
1.5 


EIGHTH    PERIOD. — EFFECTS    OF    POTASSIUM    CYANIDE    IN    INCREASED 
DOSES,  AUGUST  31  TO  SEPTEMBER  4,   igo?.48 

40 8.97  415  22  Acid                   24            492 

41 8.98  510  21  35                2 

42 8.97  500  23  46 

43 8.97  475  20  40 

44 9-03  S2©  21  50                2 

47yomited  within  the  hour  following  injection.  Some  of  the  fluid 
vomitus  ran  into  the  urine.  The  treatment  was  discontinued  and 
a  new  normal  period  inaugurated  on  the  twenty-eighth  day. 

46No  analytical  work  was  done  on  the  urine  of  this  period  because 
of  the  contamination  of  the  urine  with  fluid  vomit. 

49Vomited  during  the  night.     Urine  contained  fluid  vomit. 


21 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 
NINTH   PERIOD. — NORMAL  CONDITIONS,   SEPTEMBER   5   TO   10,    1907. 


4?  >» 

9  t  s     8 

I          B       -f.     -        .1-       '-i  «8 

&         is     !•!•>    o  111       i|  Itl 

£>c/3  f  £>0i;s  fe  bo  P-«  &S 


45 9.10             345  26                  Acid  40 

46 9.04             502  21       Faintly   alkaline  55 

47 9.06              505  21           Faintly    acid  o 

48 9.10             500  22                  Acid  23 

49 9.11              435  20                   Acid  56 

50 9.10             485  20            Amphoteric  53 

TENTH    PERIOD. — EFFECTS  OF    POTASSIUM    CYANIDE    IN  MODERATE 
DOSES,  SEPTEMBER   n   TO  15,   1907. 

51 9.20             430  20           Faintly    acid  33               2 

52 9.20             410  24            Amphoteric  64                2 

53 9.21              393  25        Faintly   alkaline  40                2 

54 9.19              513  22            Amphoteric  48                2 

55 9-21              475  23  33                2 

ELEVENTH  PERIOD. — AFTER  PERIOD,  SEPTEMBER  16  TO  20,  1907. 

56 9.18     485  22       Acid  52 

57 9-20      455  24  35 

58 9-25      420  24  29 

59 9-20      530  21  30 

60 9.20      470  21  42 


22 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

Table  XIV.— Sixth  Dog. 
Preparatory  period. 

Table  showing  the  increasing  quantities  of  meat 
fed  the  animal  daily  in  raising  it  gradually  to  a  high 
plane  of  protein  nutrition. 

(The  initial  daily  diet  was  made  up  as  follows: 
Meat,  1 08  grammes60;  cracker  meal,  36  grammes; 
lard,  27  grammes;  bone  ash,  9  grammes;  water, 
315  c.c.) 


S  *•    «  >  4-     S 

i  iJ   fl      s  &    |i 

Q  M^          >  W  Q  MS          £  M 

Oct.  12 8.10  B1i28  Oct.  19 7.95  268 

Oct.  13,., 8.09  148  Oct.  20 8.04  B3288 

Oct.  14. 8.00  168  Oct.  21 8.00  308 

Oct.  15 8.06  188  Oct.  22 8.00  328 

Oct.  1 6 8.00  208  Oct.  23 8.07  348 

Oct.   17 8.00  228  Oct.  24 8.10  M348 

Oct.  18 8.00  B2248 

The  daily  portion  of  meat  was  increased  20 
grammes  per  day  ending  with  October  23d,  on 
which  day  the  weight  of  meat  in  the  diet  was  348 
grammes.  This  weight  of  meat  was  fed  daily,  to- 
gether with  the  weights  of  the  rest  of  the  ingre- 
dients as  specified  above,  to  the  end  of  the  experi- 
ment. The  animal  was  fed  this  diet  from  October 
23d  to  November  2d  (eleven  days),  before  the 
actual  observation  of  normal  conditions  was  com- 
menced. 

50 About  13  grammes  per  kilo;  approximately  0.46  gramme  of  ni- 
trogen per  kilo. 

51  About  1 6  grammes  per  kilo;  approximately  0.56  gramme  of  ni- 
trogen per  kilo. 

62On  this  day  the  bone  ash  was  increased  to  15  grammes.  All  the 
other  ingredients  except  the  meat  remained  constant. 

53 About  32  grammes  per  kilo;  approximately  1.12  gramme  of  ni- 
trogen per  kilo. 

64  About  43  grammes  per  kilo;  approximately  1.51  gramme  of  ni- 
trogen per  kilo. 

24 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 
Table  XV.— Sixth  Dog. 

FIRST   PERIOD. — NORMAL  CONDITIONS,   NOVEMBER   3   TO   7,    1907. 


1 


3  *  *  M 

I  -4r  w  J  .  Jgu  ^.tol 

is  II"  IS  111  |||i  £il?t 

pqS  S>d  S~  £«£  D<aB 


1 8.78  395  25  Acid.                0.2747  33 

2 8.71  448  24  0.2871  53 

3.......  8.73  413  26  0.2751  43 

4 8.74  503  23  0.3400  ii 

5 8.80  390  26  0.3076  41 

SECOND    PERIOD. — EFFECTS    OF    POTASSIUM    CYANIDE,    NOVEMBER    8 
TO  13,  1907. 

6 8.80  467  25  Acid.               0.3147  26      0.5 

7 8.81  432  25  0.3394  3i       i 

8 8.95  355  26  0.2594  15      1.5 

9 8.92  423  24  0.4495  33      1.75 

10 9.00  302  27  0.2448  34      2 

" 8.96  457  25  0.3254  35      2.5 


THIRD  PERIOD. — AFTER   PERIOD,   NOVEMBER    14  TO   20,  1907. 

12 8.94         470  23  Acid.               0.3141  19 

13 9-co         365  28                                       0.3119  20 

14 9-09         4i5  26                                       0.3433  26 

IS 9-20         275  29                                       0.1913  29 

16 9.20         380  28                                       0.3768  37 

17 9-09          572  23                                         0.4637  37 

18 9.21          339  23                                         0.3306  23 

Discussion  of  the  chemical  results  of  the  experi- 
ments with  animals  on  high  planes  of  protein 
nutrition  (fifth  and  sixth  dogs,  Tables  XII- 
XVI). 

In  both  of  these  experiments  the  average  daily 
total  elimination  of  urinary  nitrogen  was  dimin- 
ished during  the  periods  of  cyanide  injection.  In 
the  experiment  on  the  fifth  dog  the  ammonia  ni- 
trogen rose  markedly  during  the  period  (VII)  in 
which  small  doses  of  cyanide  were  administered 
(Tables  XII-XIII),  and  increased  slightly  during 
the  period  (X)  in  which  moderate  doses  of  cyanide 
were  injected.  There  was  a  similar  slightly  in- 
creased elimination  of  ammonia  nitrogen  during  the 
cyanide  period  in  the  experiment  on  the  sixth  dog 
(Tables  XV-XVI). 

25 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 


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26 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

In  the  experiment  on  the  fifth  dog,  the  urea  ni- 
trogen fell  slightly  during  the  period  (VII)  in 
which  relatively  small  doses  of  cyanide  were  inject- 
ed, and  also  during  the  period  in  which  compara- 
tively moderate  doses  of  cyanide  (X)  were  admin- 
istered. On  the  basis  of  its  proportion  of  the  total 
nitrogen,  the  urea  nitrogen  in  the  urine  of  the  fifth 
dog  fell  slightly  in  the  first  of  the  two  periods  re- 
ferred to,  but  rose  in  the  latter  period.  During  the 
cyanide  period  of  the  experiment  on  the  sixth  dog 
(Tables  XV-XVI)  the  urea  nitrogen  rose  in 
quantity. 

The  allantoin  nitrogen  was  diminished  after  ad- 
ministration of  comparatively  small  doses  of  cyan- 
ide in  the  fifth  dog,  but  was  increased  after  mod- 
erate doses  in  the  same  animal.  The  allantoin  ni- 
trogen was  diminished  during  the  cyanide  period 
of  the  experiment  on  the  sixth  dog. 

In  the  experiment  on  the  fifth  dog  the  uric  acid 
nitrogen  was  fairly  constant  throughout,  but  in  the 
case  of  the  sixth  dog  the  eliminated  amount  in  the 
cyanide  period  was  larger  than  that  of  the  first 
period,  while  the  excreted  quantity  in  the  after 
period  was  greater  than  the  combined  amounts  of 
uric  acid  nitrogen  in  the  fore  period  and  the  cyan- 
ide period. 

In  both  experiments,  creatinin  nitrogen  appeared 
to  be  diminished  by  the  cyanide  treatment. 

The  "undetermined  nitrogen"  was  but  little  af- 
fected by  small  doses  of  cyanide  in  the  fifth  dog, 
but  was  decreased  markedly  in  the  same  animal 
after  treatment  with  moderate  doses  of  cyanide. 
The  same  decrease  in  undetermined  nitrogen  ap- 
peared after  the  cyanide  treatment  of  the  sixth 
dog. 

Histological  data — Fifth  and  sixth  dogs. 

The  microscopical  examination  of  the  viscera  of 
these  dogs  showed  the.  following: 

Fifth  dog.     Kidney — There  was  marked  conges- 

27 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

tion,  with  well  marked  albuminous  degeneration  of 
the  epithelium  of  the  convoluted  tubules.  The 
epithelium  was  swollen  and  in  places  disintegrating. 
The  lumina  of  the  tubules  contained  a  large  amount 
of  granular  matter.  The  nuclei  were  intact.  Some 
glomeruli  were  much  shrunken  and  were  composed 
almost  entirely  of  connective  tissue  cells.  In  the 
neighborhood  of  some  glomeruli  and  along  the 
course  of  some  blood  vessels  were  large  areas, 
rather  sharply  circumscribed,  of  small  round  cell 
proliferation. 

Liver. — This  was  the  seat  of  mild  parenchymatous 
degeneration,  slightly  more  marked  about  the  cen- 
tral vein  than  in  the  periphery  of  the  lobule.  The 
degeneration,  which  was  albuminous  in  nature,  was 
slight  and  accompanied  by  very  moderate  vacuoli- 
zation  of  the  cells.  The  congestion  was  not  marked. 

Heart — Normal.  Spleen — Normal.  Pancreas — 
Normal. 

Sixth  dog.  Kidney — The  cortex  was  much  con- 
gested and  the  tubular  epithelium  was  the  seat  of 
marked  albuminous  degeneration,  which  was  most 
evident  in  the  convoluted  tubules.  The  cells  were 
greatly  swollen  and  granular,  and  irregular  in  out- 
line. The  lumina  contained  granular  matter.  No 
distinct  proliferation  of  connective  tissue  could  be 
made  out.  The  glomeruli  were  normal. 

Liver — There  was  mild  albuminous  degeneration 
of  the  liver  cells,  slightly  more  marked  about  the 
central  vein  than  in  the  periphery  of  the  lobule. 
There  was  very  slight  vacuolization  of  the  cells. 
There  was  no  congestion. 

Heart — Normal.  Spleen — Normal.  Pancreas — 
Normal. 

Historical. 

The  first  investigation  of  the  influence  of  potas- 
sium cyanide  upon  nitrogenous  metabolism  was 
made  by  Loewy."  In  a  preliminary  communica- 

"Loewy:   Centralblatt  fur  Physiologic,   xix,   p.   856,    1906. 
28 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

tion  he  stated  that  on  the  day  of  poisoning,  and  on 
the  day  after,  the  nitrogen  elimination  was  marked- 
ly increased.  The  output  of  amino-acid  nitrogen 
also  seemed  to  be  decidedly  increased. 

In  a  second  paper  on  this  subject,  Loewy86  stated 
that  the  total  quantity  of  excreted  nitrogen  did  not 
increase  in  all  cases.  He  concluded  that  the  plane 
of  nutrition  has  some  relation  to  the  manner  in 
which  cyanide  affects  nitrogenous  metabolism. 
That  conclusion  is  in  accord  with  the  prior  results 
obtained  by  Frankel  and  Geppert57  in  their  re- 
searches on  diminished  oxygen  supply.  They  es- 
tablished a  similar  relation  between  increased  ni- 
trogen elimination  arid  the  plane  of  nutrition.  The 
increase  of  excreted  nitrogen  was  always  least  in 
fasting  animals,  and  invariably  appeared  later. 

As  the  first  series  of  experiments  in  this  study 
were  nearing  completion,  we  were  informed  that  a 
similar  investigation  was  being  conducted  by 
Richards  and  Wallace.58  Their  published  results 
do  not  show  any  marked  difference  in  the  effects 
of  cyanide  on  two  dogs,  on  different  planes  of  nu- 
trition. Richards  and  Wallace  observed  an  in- 
crease in  the  total  of  excreted  nitrogen  and  urea 
nitrogen,  with  no  increase  of  ammonia  nitrogen  in 
the  urine.  Creatinin  was  markedly  diminished  in 
the  urine  on  the  days  of  severe  poisoning.  The  un- 
determined nitrogen  increased  in  the  urine  on  the 
days  of  severe  poisoning. 

Conclusions. 

Potassium  cyanide,  injected  subcutaneously  in 
gradually  increasing  daily  amounts  from  one  half 
to  three  and  a  half  milligrammes  per  kilo,  through 
periods  ranging  from  four  to  twelve  days,  failed  to 
cause  any  marked  increase  in  the  total  elimination 
of  nitrogen  in  the  urine.  In  six  out  of  the  nine 

58Loewy:  Biochemische  Zeitschrift,  iii,  p.  439,   1907. 
"Frankel  and  Geppert.     Cited  by  Loewy,  loc.  sit. 
^Richards    and    Wallace:    Journal   of   Biological   Chemistry,    iv,    p. 
179,   1908. 

2Q 


Welker  and  Ditman:  Biochemical  and  Anatomical  Changes. 

cyanide  periods,  however,  the  total  amount  of  ni- 
trogen excreted  in  the  urine  was  actually  dimin- 
ished. In  the  case  of  the  three  other  periods  the 
increase  observed  was  very  slight. 

Possibly  the  urinary  excretion  of  nitrogen  was  not 
materially  increased  in  these  experiments,  after  the 
cyanide  injections,  because  the  doses  were  too 
small  to  produce  convulsions  or  to  cause  the  asso- 
ciated conditions  which  may  markedly  augment 
catabolism,  and  which  may  have  been  largely  re- 
sponsible for  the  increased  elimination  of  nitrogen 
that  was  observed  by  some  of  our  predecessors  in 
such  studies. 

The  plane  of  protein  nutrition  seems  to  influ- 
ence the  degree  to  which  subcutaneous  injections  of 
cyanide  affect  the  elimination  of  some  of  the  forms 
of  nitrogen  in  the  urine.  A  comparison  of  the  re- 
sults for  urea  and  undetermined  nitrogen  in  the 
urines  of  the  animals  on  high  and  low  planes  of 
protein  nutrition  makes  it  evident  that  the  percent- 
age of  urea  nitrogen  increased  and  that  of  the 
undetermined  products  decreased  in  the  urines  from 
the  dogs  on  high  protein  nutrition  planes,  whereas 
the  reverse  in  each  case  was  noted  for  the  urines 
from  the  dogs  on  low  protein  nutrition  planes. 

Further  work  will  be  necessary  before  the  open 
questions  that  remain  can  be  answered  with  assur- 
ance. 

We  wish  to  express  our  most  heartfelt  thanks  to 
Professor  William  J.  Gies  for  his  kind  advice  and 
guidance  in  the  conduct  of  these  experiments. 


BIOGRAPHICAL 

William  Henry  Welker  received  his  early  education  in  the 
public  schools  and  prepared  for  college  at  Perkiomen  Seminary 
and  Ursinus  College.  He  taught  public  school  at  Klinesville,  Pa., 
during  the  terms  of  1898-99  and  1899-1900.  He  graduated  from 
Perkiomen  Seminary  in  the  spring  of  1900,  entered  Lehigh 
University  in  the  fall  of  1900,  and  graduated  from  Lehigh  in  1904, 
receiving  the  degree  of  Analytical  Chemist.  He  was  undergradu- 
ate instructor  in  general  chemistr}7  at  Lehigh  in  1902-03  and  1903- 
04,  and  undergraduate  instructor  in  physics  in  1903-04.  He  was 
appointed  Assistant  in  Chemistry  at  Lehigh  University  at  the  time 
of  his  graduation  in  1904,  but  resigned  during  the  summer  of  1904 
to  accept  the  position  of  Assistant  in  Physiological  Chemistry  in 
Columbia  University,  at  the  College  of  Physicians  and  Surgeons. 
He  served  in  this  capacity  until  Dec.  i,  1906,  when,  in  addition, 
he  became  Pathological  Assistant  in  Obstetrics  at  the  College  of 
Physicians  and  Surgeons.  Recently  he  resigned  both  these 
positions  to  accept  that  of  Demonstrator  of  Physiological  Chemistry 
at  the  University  of  Pennsylvania. 

PUBLICATIONS 

Experiments  to  determine  the  influence  of  the  bromides  of 
barium  and  radium  on  protein  metabolism.  (With  William  N. 
Berg).  Journal  of  Biological  Chemistry,  1906,  i,  p.  371. 

A  contribution  to  our  knowledge  of  the  effects  of  urinary  pre- 
servatives on  urinary  analysis.  New  York  Medical  Journal,  1907 
(September  21),  p.  552. 

A  simple  electrical  annunciator  for  use  in  metabolism  experi- 
ments and  in  connection  with  filtration,  distillation  and  similar 
operations.  American  Journal  of  Physiology,  1907,  xx,  p.  358. 

On  some  biochemical  and  anatomical  changes  induced  in  dogs 
by  potassium  cyanid.  (With  Norman  E.  Ditman).  New  York 
Medical  Journal,  1908  (July  n),  p.  59. 


UNTVEESITY   OF   CALIFOENIA   LIBEAEY, 
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